New aspects of a small GTPase RAB35 in brain development and function

In eukaryotic cells,organelles in the secretory,lysosomal,and endocytic pathways actively exchange biological materials with each other through intracellular membrane trafficking,which is the process of transporting the cargo of proteins,lipids,and other molecules to appropriate compartments via transport vesicles or intermediates.These processes are strictly regulated by various small GTPases such as the RAS-like in rat brain(RAB)protein family,which is the largest subfamily of the RAS superfamily.Dysfunction of membrane trafficking affects tissue homeostasis and leads to a wide range of diseases,including neurological disorders and neurodegenerative diseases.Therefore,it is important to understand the physiological and pathological roles of RAB proteins in brain function.RAB35,a member of the RAB family,is an evolutionarily conserved protein in metazoans.A wide range of studies using cultured mammalian cells and model organisms have revealed that RAB35 mediates various processes such as cytokinesis,endocytic recycling,actin bundling,and cell migration.RAB35 is also involved in neurite outgrowth and turnover of synaptic vesicles.We generated brain-specific Rab35 knockout mice to study the physiological roles of RAB35 in brain development and function.These mice exhibited defects in anxiety-related behaviors and spatial memory.Strikingly,RAB35 is required for the precise positioning of pyramidal neurons during hippocampal development,and thereby for normal hippocampal lamination.In contrast,layer formation in the cerebral cortex occurred superficially,even in the absence of RAB35,suggesting a predominant role for RAB35 in hippocampal development rather than in cerebral cortex development.Recent studies have suggested an association between RAB35 and neurodegenerative diseases,including Parkinson's disease and Alzheimer's disease.In this review,we provide an overview of the current understanding of subcellular functions of RAB35.We also provide insights into the physiological role of RAB35 in mammalian brain development and function,and discuss the involvement of RAB35 dysfunction in neurodegenerative diseases.

Establishment of human cerebral organoid systems to model early neural development and assess the central neurotoxicity of environmental toxins

Human brain development is a complex process,and animal models often have significant limitations.To address this,researchers have developed pluripotent stem cell-derived three-dimensional structures,known as brain-like organoids,to more accurately model early human brain development and disease.To enable more consistent and intuitive reproduction of early brain development,in this study,we incorporated forebrain organoid culture technology into the traditional unguided method of brain organoid culture.This involved embedding organoids in matrigel for only 7 days during the rapid expansion phase of the neural epithelium and then removing them from the matrigel for further cultivation,resulting in a new type of human brain organoid system.This cerebral organoid system replicated the temporospatial characteristics of early human brain development,including neuroepithelium derivation,neural progenitor cell production and maintenance,neuron differentiation and migration,and cortical layer patterning and formation,providing more consistent and reproducible organoids for developmental modeling and toxicology testing.As a proof of concept,we applied the heavy metal cadmium to this newly improved organoid system to test whether it could be used to evaluate the neurotoxicity of environmental toxins.Brain organoids exposed to cadmium for 7 or 14 days manifested severe damage and abnormalities in their neurodevelopmental patterns,including bursts of cortical cell death and premature differentiation.Cadmium exposure caused progressive depletion of neural progenitor cells and loss of organoid integrity,accompanied by compensatory cell proliferation at ectopic locations.The convenience,flexibility,and controllability of this newly developed organoid platform make it a powerful and affordable alternative to animal models for use in neurodevelopmental,neurological,and neurotoxicological studies.

Postnatal development of rat retina:a continuous observation and comparison between the organotypic retinal explant model and in vivo development

The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures.However,the lack of systematic and continuous comparison between in vivo retinal development and the organotypic retinal explant culture makes this model controversial in postnatal retinal development studies.Thus,we aimed to verify the feasibility of using this model for postnatal retinal development studies by comparing it with the in vivo retina.In this study,we showed that postnatal retinal explants undergo normal development,and exhibit a consistent structure and timeline with retinas in vivo.Initially,we used SOX2 and PAX6 immunostaining to identify retinal progenitor cells.We then examined cell proliferation and migration by immunostaining with Ki-67 and doublecortin,respectively.Ki-67-and doublecortin-positive cells decreased in both in vivo and explants during postnatal retinogenesis,and exhibited a high degree of similarity in abundance and distribution between groups.Additionally,we used Ceh-10 homeodomain-containing homolog,glutamate-ammonia ligase(glutamine synthetase),neuronal nuclei,and ionized calcium-binding adapter molecule 1 immunostaining to examine the emergence of bipolar cells,Müller glia,mature neurons,and microglia,respectively.The timing and spatial patterns of the emergence of these cell types were remarkably consistent between in vivo and explant retinas.Our study showed that the organotypic retinal explant culture model had a high degree of consistency with the progression of in vivo early postnatal retina development.The findings confirm the accuracy and credibility of this model and support its use for long-term,systematic,and continuous observation.

"Three-in-One" Ecological Development and Efficient Utilization Model of Aggregates

Green mining and the formation of an effective and efficient development model have become key issues that aggregates enterprises around the world need to solve urgently.On the basis of analyzing the development status of aggregates industry in Xiluodu area,the paper studied the main problems faced in the construction of green aggregates mines at present,and proposed a"three-in-one"ecological,intelligent and efficient green mine construction model for"ecological development","green logistics"and"solid waste recycling"of aggregates.The study has certain theoretical value and practical significance for the construction of green aggregates mine in Xiluodu area.

Knowledge mapping of research on sustainable utilization of landscape resources:a case study of Guilin National Innovation Demonstration Zone for sustainable development in China

Worldwide interest has increasingly focused on the sustainable utilization of landscape as a resource in urban areas,emphasizing its ecological,cultural and social significance.This study examines Guilin City,China,as a representative case study due to its rich landscape resources and status as a national innovation demonstration zone for implementing the 2030 Agenda for Sustainable Development.This study uses bibliometric visualization tools like CiteSpace and VOSviewer to analyze research trends from 1980 to 2021 in the Chinese Academic Journal Network Publishing Database(CNKI).The results show increasing academic interest over three stages:initiation(1982-1997),exploration(1998-2004),and diversified development(2005-2021).Contributions are predominantly from local academic and tourism sectors,indicating a strong regional influence;however,relatively weak interinstitutional collaboration occurs,suggesting potential for more integrated research efforts.Primary research is also concentrated within economic disciplines,particularly tourism-related ones.The evolution of research frontiers reveals three main paths:urban development strategies,industrial economic theories and empirical validation,and ecosystem analysis and evaluation.A multidisciplinary approach and stronger collaborative efforts are crucial to enhance research on ecological values and empirical models while supporting evidence-based urban development strategies in Guilin City and comparable cities globally.

Study on Utilization and Development of Hot Spring in Liaocheng City

The basic situation and formation mechanism of hot spring in Liaocheng were introduced,and its utilization value as well as exploitation and utilization status were also analyzed in this study,which provided strategies for exploitation and utilization of hot spring in Liaocheng.

Research Progress on the Development and Utilization of Proteins in Edible Insects

The recent progress on the proteins in edible insects was summarized, in- cluding the nutritional value, healthy value, food safety, extraction and content mea- surement of the proteins in edible insects, as well as the main development direc- tion of edible insect proteins. And we also pointed out the bottlenecks that restricted the development and utilization of proteins in edible insects, and proposed the prospect of the research, development and utilization of edible insects proteins.

Rural Biomass Energy Utilization and Sustainable Developmental Strategies in Tibet

[Objective] This study was to provide theoretical basis for getting sustainable development of rural energy in Tibet into reality.[Method] By reviewing the rural energy resources in Tibet,we analyzed the characteristics and potential of rural biomass utilization in Tibet,and further put forward the sustainable countermeasures on the firewood substitution in Tibet.[Results] Renewable energies including hydraulic,geothermal,solar and wind resources are abundant in Tibet,while there is just a few of fossil energy resources such as oil and coal,with uneven distribution and poor exploration conditions.Traditional consumption of biomass energy resource accounts for a large proportion of the total energy consumption in Tibetan rural districts,which causes potential damage to the fragile ecological environment on the Tibetan Plateau.The excessive use of biomass energy destroyed the vegetation and evoked the environment deterioration such as the intensification of the water and soil loss and the declining of the soil fertility.[Conclusion] It is essential for Tibet to change its rural energy consumption structure,implement the Tibetan firewood alternative energy strategy and try to make full use of renewable energy such as solar energy,wind energy,hydro-energy instead of native vegetation and animal＇s droppings in order to reduce the adverse impacts of the irrational energy consumption on the ecological environment in Tibet.

Development and Utilization of Naturally Colored Cocoon Resources of Bombyx mori

This paper summarized the diversity and prominent characteristics of naturally colored cocoon resources of Bombyx mori, and introduced the production methods of colored cocoons, the development and utilization of naturally colored cocoon resources at home and abroad as well as the variety breeding instances, based on which suggestions were proposed for the development of colored cocoon industry.

Distribution, Development and Utilization of Bioactive Components Resources of Russian Melilotus officinalis

To confirm the content and distribution of bioactive components in roots, stems, leaves and grains of Russian Me/i/otus officina/is ＇Sligen 1 ＂, an experiment involving bioactive components detection and field planting was conducted. Kjeldahl method and high performance gas chromatography were used to detect the protein. The automatic amino acid analyzer was used to detect amino acid. Gas chromatography-mass spectrometry （GC-MS） and molecular distillation apparatus were used to confirm the fatty acid content. Gravimetric method was used to detect saponin and alkaloid contents. High performance liquid chromatography was used to detect the coumarin content. The polyphenol content was determined by forint reagent colorimetric method. Anthracene copper-sulfuric acid colorimetry was used to detect the polysaccharide, and aluminum nitrate-sodium nitrite-sodium hydroxide col- orimetric method was used to confirm flavone. The results showed that protein and amino acid were mainly distributed in leaves and seeds. The fatty acid was mainly distributed in seeds. The saponin was mainly distributed in roots and leave. The polyphenol was mainly distributed in roots, stems and leaves. The alkaloid was mainly distributed in stems, leaves and seeds. Seeds mainly contained coumarin. M. officinalis had little polysaccharide and flavone. The total production of bioactive in- gredients was 3 016.3 kg/hm2. The output of crude protein, amino acids and fatty acids was 1 400.6, 1 246.8, 112.2, respectively. The yields of effective components saponins, alkaloids, polyphenols and coumarin were 97.0, 77.9, 41.7, 40.1 kg/hm2, respectively. After separation, extraction and purification, those components can be used as the natural ingredients to develop medicine, food, chemical and other biotechnological products.

Development and Utilization of the World＇s and China＇s Bulk Mineral Resources and their Supply and Demand Situation in the Next Twenty Years

Bulk mineral resources of iron ores, copper ores, bauxite, lead ores, zinc ores and potassium salt play a pivotal role on the world＇s and China＇s economic development. This study analyzed and predicted their resources base and potential, development and utilization and their world＇s and China＇s supply and demand situation in the future 20 years. The supply and demand of these six bulk mineral products are generally balanced, with a slight surplus, which will guarantee the stability of the international mineral commodity market supply. The six mineral resources （especially iron ores and copper ores） are abundant and have a great potential, and their development and utilization scale will gradually increase. Till the end of 2014, the reserve- production ratio of iron, copper, bauxite, lead, zinc ores and potassium salt was 95 years, 42 years, 100 years, 17 years, 37 years and 170 years, respectively. Except lead ores, the other five types all have reserve-production ratio exceeding 20 years, indicative of a high resources guarantee degree. If the utilization of recycled metals is counted in, the supply of the world＇s six mineral products will exceed the demand in the future twenty years. In 2015-2035, the supply of iron ores, refined copper, primary aluminum, refined lead, zinc and potassium salt will exceed their demand by 0.4-0.7 billion tons （Gt）, 5.0-6.0 million tons （Mt）, 1.1-8.9 Mt, 1.0-2.0 Mt, 1.2-2.0 Mt and 4.8-5.6 Mt, respectively. It is predicted that there is no problem with the supply side of bulk mineral products such as iron ores, but local or structural shortage may occur because of geopolitics, monopoly control, resources nationalism and trade friction. Affected by China＇s compressed industrialized development model, the demand of iron ores （crude steel）, potassium salt, refined lead, refined copper, bauxite （primary aluminum） and zinc will gradually reach their peak in advance. The demand peak of iron ores （crude steel） will reach around 2015, 2016 for potassium salt, 2020 for refined lead, 2021 for bauxite （primary aluminum）, 2022 for refined copper and 2023 for zinc. China＇s demand for iron ores （crude steel）, bauxite （primary aluminum） and zinc in the future 20 years will decline among the world＇s demand, while that for refined copper, refined lead and potassium salt will slightly increase. The demand for bulk mineral products still remains high. In 2015-2035, China＇s accumulative demand for iron ores （crude steel） will be 20.313 Gt （13.429 Gt）, 0.304 Gt for refined copper, 2.466 Gt （0.616 Gt） of bauxite （primary aluminum）, 0.102 Gt of refined lead, 0.138 Gt of zinc and 0.157 Gt of potassium salt, and they account for the world＇s YOY （YOY） accumulative demand of 35.17%, 51.09%, 48.47%, 46.62%, 43.95% and 21.84%, respectively. This proportion is 49.40%, 102.52%, 87.44%, 105.65%, 93.62% and 106.49% of that in 2014, respectively. From the supply side of China＇s bulk mineral resources, it is forecasted that the accumulative supply of primary （mine） mineral products in 2015-2035 is 4.046 Gt of iron ores, 0.591 Gt of copper, 1.129 Gt of bauxite, 63.661 Mt of （mine） lead, 0.109 Gt of （mine） zinc and 0.128 Gt of potassium salt, which accounts for 8.82%, 13.92%, 26.67%, 47.09%, 33.04% and 15.56% of the world＇s predicted YOY production, respectively. With the rapid increase in the smelting capacity of iron and steel and alumina, the rate of capacity utilization for crude steel, refined copper, alumina, primary aluminum and refined lead in 2014 was 72.13%, 83.63%, 74.45%, 70.76% and 72.22%, respectively. During 2000-2014, the rate of capacity utilization for China＇s crude steel and refined copper showed a generally fluctuating decrease, which leads to an insufficient supply of primary mineral products. It is forecasted that the supply insufficiency of iron ores in 2015-2035 is 17.44 Gt, 0.245 Gt of copper in copper concentrates, 1.337 Gt of bauxite, 38.44 Mt of lead in lead concentrates and 29.19 Mt of zinc in zinc concentrates. China has gradually raised the utilization of recycled metals, which has mitigated the insufficient supply of primary metal products to some extent. It is forecasted that in 2015-2035 the accumulative utilization amount of steel scrap （iron ores） is 3.27 Gt （5.08 Gt）, 70.312 Mt of recycled copper, 0.2 Gt of recycled aluminum, 48 Mt of recycled lead and 7.7 Mt of recycled zinc. The analysis on the supply and demand situation of China＇s bulk mineral resources in 2015-2035 suggests that the supply-demand contradiction for these six types of mineral products will decrease, indicative of a generally declining external dependency. If the use of recycled metal amount is counted in, the external dependency of China＇s iron, copper, bauxite, lead, zinc and potassium salt will be 79%, 65%, 26%, 8%, 16% and 18% in 2014, respectively. It is predicted that this external dependency will decrease to 62%, 64%, 20%, -0.93%, 16% and 14% in 2020, respectively, showing an overall decreasing trend. We propose the following suggestions correspondingly. （1） The demand peak of China＇s crude steel and potassium salt will reach during 2015-2023 in succession. Mining transformation should be planned and deployed in advance to deal with the arrival of this demand peak. （2） The supply-demand contradiction of China＇s bulk mineral resources will mitigate in the future 20 years, and the external dependency will decrease accordingly. It is suggested to adjust the mineral resources management policies according to different minerals and regions, and regulate the exploration and development activities. （3） China should further establish and improve the forced mechanism of resolving the smelting overcapacity of steel, refined copper, primary aluminum, lead and zinc to really achieve the goal of ＂reducing excess production capacity＂. （4） In accordance with the national strategic deployment of ＂One Belt One Road＂, China should encourage the excess capacity of steel, copper, alumina and primary aluminum enterprises to transfer to those countries or areas with abundant resources, high energy matching degree and relatively excellent infrastructure. Based on the national conditions, mining condition and geopolitics of the resources countries, we will gradually build steel, copper, aluminum and lead-zinc smelting bases, and potash processing and production bases, which will promote the excess capacity to transfer to the overseas orderly. （5） It is proposed to strengthen the planning and management of renewable resources recycling and to construct industrial base of renewable metal recycling. （6） China should promote the comprehensive development and utilization of paragenetic and associated mineral species to further improve the comprehensive utilization of bulk mineral resources.

Research on the Utilization and Development of the Ring Park in Hefei City of China

To disclose problems of the Ring Park in Hefei City (the park around the inner city of Hefei, Anhui Province), propose reasonable renovation plans and discuss its future development trend, this study investigates the park through field investigation, questionnaire survey and interview. Through exploring development background and history of the park, it is proposed that the park has no sufficient service facilities but potential dangers, some of its vegetation is destroyed and water quality is greatly reduced, then comprehensive solutions are given as:to improve service facilities by organizing pedestrian and traffic flow in the park and setting the city ring buses; to reduce potential safety hazards by enhancing crime control and daily safety control within the park; to improve the overall eco-environment of the park by renovating its water and embankment, maintaining and adjusting vegetations. Prospects of regional tourism development and conceptions for the further development of this park are proposed, and the authors suggest that more scenic spots should be created and tourist routes reasonably organized.

Development and Utilization of Saline Lake Resources and Protection of Ecological Environment in Qaidam Basin in Qinghai Province

Qaidam basin is an important part of National Key Development Area–Lanzhou-Xining Area,is a key region of resource development in Western Development,is a National Circular Economy Pilot Zone.The focus of

Structural properties of residual carbon in coal gasification fine slag and their influence on flotation separation and resource utilization:A review

Coal gasification fine slag(FS)is a typical solid waste generated in coal gasification.Its current disposal methods of stockpil-ing and landfilling have caused serious soil and ecological hazards.Separation recovery and the high-value utilization of residual carbon(RC)in FS are the keys to realizing the win-win situation of the coal chemical industry in terms of economic and environmental benefits.The structural properties,such as pore,surface functional group,and microcrystalline structures,of RC in FS(FS-RC)not only affect the flotation recovery efficiency of FS-RC but also form the basis for the high-value utilization of FS-RC.In this paper,the characteristics of FS-RC in terms of pore structure,surface functional groups,and microcrystalline structure are sorted out in accordance with gasification type and FS particle size.The reasons for the formation of the special structural properties of FS-RC are analyzed,and their influence on the flotation separation and high-value utilization of FS-RC is summarized.Separation methods based on the pore structural characterist-ics of FS-RC,such as ultrasonic pretreatment-pore-blocking flotation and pore breaking-flocculation flotation,are proposed to be the key development technologies for improving FS-RC recovery in the future.The design of low-cost,low-dose collectors containing polar bonds based on the surface and microcrystalline structures of FS-RC is proposed to be an important breakthrough point for strengthening the flotation efficiency of FS-RC in the future.The high-value utilization of FS should be based on the physicochemical structural proper-ties of FS-RC and should focus on the environmental impact of hazardous elements and the recyclability of chemical waste liquid to es-tablish an environmentally friendly utilization method.This review is of great theoretical importance for the comprehensive understand-ing of the unique structural properties of FS-RC,the breakthrough of the technological bottleneck in the efficient flotation separation of FS,and the expansion of the field of the high value-added utilization of FS-RC.

Harmonious Development of Utilization and Protection of Tidal Flats and Wetlands-A Case Study in Shanghai Area

Spatial, environmental, and material resources are important for tidal flats and wetlands, thus, a harmonious balance between utilization and protection should be maintained. In the present study, the concept of tidal flat is defined, and its spatial limitation is also clarified. Located at the Yangtze Estuary, tidal flats in Shanghai area are selected for study. The relationship between water discharge and sediment transport of the Yangtze River, allowed exploration intensity and needs for wetland protection are discussed in the paper. The dynamic process and changing trend for different types of tidal flats are illustrated in detail. Regulations are proposed for a reasonable exploitation, dynamic protection, and ecosystem restoration of each individual tidal fiat and wetland. Some valuable suggestions are put forward for further study about the sustain＇able exploitation of offshore resources in Shanghai area.

Priority-sequence of mineral resources' development and utilization based on grey relational analysis method

Generally, the sequence decision of the development and utilization of Chinese mineral resources is based on national and provincial overall plan of the mineral resources. Such plan usually cannot reflect the relative size of the suitability of the development and utilization of mineral resources. To solve the problem, the paper has selected the gift condition, the market condition, the technological condition,socio-economic condition and environmental condition as the starting-points to analyze the influential factors of the priority-sequence of mineral resources' development and utilization. The above 5 conditions are further specified into 9 evaluative indicators to establish an evaluation indicator system. At last,we propose a decision model of the priority sequence based on grey relational analysis method, and figure out the observation objects by the suitability index of development. Finally, the mineral resources of a certain province in China were analyzed as an example. The calculation results indicate that silver(2.0057), coal(1.9955), zinc(1.9442), cement limestone(1.9077), solvent limestone(1.5624) and other minerals in the province are suitable for development and utilization.

Conventional Geothermal Systems and Unconventional Geothermal Developments: An Overview

This paper provides an overview of conventional geothermal systems and unconventional geothermal developments as a common reference is needed for discussions between energy professionals. Conventional geothermal systems have the heat, permeability and fluid, requiring only drilling down to °C, normal heat flow or decaying radiogenic granite as heat sources, and used in district heating. Medium-temperature (MT) 100°C - 190°C, and high-temperature (HT) 190°C - 374°C resources are mostly at plate boundaries, with volcanic intrusive heat source, used mostly for electricity generation. Single well capacities are °C - 500°C) and a range of depths (1 m to 20 Km), but lack permeability or fluid, thus requiring stimulations for heat extraction by conduction. HVAC is 1 - 2 m deep and shallow geothermal down to 500 m in wells, both capturing °C, with °C are either advanced by geothermal developers at <7 Km depth (Enhanced Geothermal Systems (EGS), drilling below brittle-ductile transition zones and under geothermal fields), or by the Oil & Gas industry (Advanced Geothermal Systems, heat recovery from hydrocarbon wells or reservoirs, Superhot Rock Geothermal, and millimeter-wave drilling down to 20 Km). Their primary aim is electricity generation, relying on closed-loops, but EGS uses fractures for heat exchange with earthquake risks during fracking. Unconventional approaches could be everywhere, with shallow geothermal already functional. The deeper and hotter unconventional alternatives are still experimental, overcoming costs and technological challenges to become fully commercial. Meanwhile, the conventional geothermal resources remain the most proven opportunities for investments and development.

Development and technology status of energy storage in depleted gas reservoirs

Utilizing energy storage in depleted oil and gas reservoirs can improve productivity while reducing power costs and is one of the best ways to achieve synergistic development of"Carbon Peak–Carbon Neutral"and"Underground Resource Utiliza-tion".Starting from the development of Compressed Air Energy Storage(CAES)technology,the site selection of CAES in depleted gas and oil reservoirs,the evolution mechanism of reservoir dynamic sealing,and the high-flow CAES and injection technology are summarized.It focuses on analyzing the characteristics,key equipment,reservoir construction,application scenarios and cost analysis of CAES projects,and sorting out the technical key points and existing difficulties.The devel-opment trend of CAES technology is proposed,and the future development path is scrutinized to provide reference for the research of CAES projects in depleted oil and gas reservoirs.

Development and Utilization of Geological Tourism Resources in Xiling Gorge of the Three Gorges

To comprehensively and reasonably develop and utilize geological tourism resources in Xiling Gorge(one of the Three Gorges),this study through classifying geological tourism resources in this section combined its geological formation,strata relationship,distribution of natural and cultural tourist resources,comprehensively analyzed problems in the present development,and potentials of local geo-tourist resources.Besides,a feasible work plan on the development of geological tourism resources along the Yangtze River,south of the River and north of the River was designed to present an optimal way for the development of local geo-tourism resources,in order to promote the sound development of geological tourism and other tourism industries in this region.

BMPRⅡ^(+)neural precursor cells isolated and characterized from organotypic neurospheres:an in vitro model of human fetal spinal cord development

Roof plate secretion of bone morphogenetic proteins(BMPs)directs the cellular fate of sensory neurons during spinal cord development,including the formation of the ascending sensory columns,though their biology is not well understood.Type-ⅡBMP receptor(BMPRⅡ),the cognate receptor,is expressed by neural precursor cells during embryogenesis;however,an in vitro method of enriching BMPRⅡ^(+)human neural precursor cells(hNPCs)from the fetal spinal cord is absent.Immunofluorescence was undertaken on intact second-trimester human fetal spinal cord using antibodies to BMPRⅡand leukemia inhibitory factor(LIF).Regions of highest BMPRⅡ^(+)immunofluorescence localized to sensory columns.Parenchymal and meningeal-associated BMPRⅡ^(+)vascular cells were identified in both intact fetal spinal cord and cortex by co-positivity with vascular lineage markers,CD34/CD39.LIF immunostaining identified a population of somas concentrated in dorsal and ventral horn interneurons,mirroring the expression of LIF receptor/CD118.A combination of LIF supplementation and high-density culture maintained culture growth beyond 10 passages,while synergistically increasing the proportion of neurospheres with a stratified,cytoarchitecture.These neurospheres were characterized by BMPRⅡ^(+)/MAP2ab^(+/–)/βⅢ-tubulin^(+)/nestin^(–)/vimentin^(–)/GFAP^(–)/NeuN^(–)surface hNPCs surrounding a heterogeneous core ofβⅢ-tubulin^(+)/nestin^(+)/vimentin^(+)/GFAP^(+)/MAP2ab^(–)/NeuN^(–)multipotent precursors.Dissociated cultures from tripotential neurospheres contained neuronal(βⅢ-tubulin^(+)),astrocytic(GFAP+),and oligodendrocytic(O4+)lineage cells.Fluorescence-activated cell sorting-sorted BMPRⅡ^(+)hNPCs were MAP2ab^(+/–)/βⅢ-tubulin^(+)/GFAP^(–)/O4^(–)in culture.This is the first isolation of BMPRⅡ^(+)hNPCs identified and characterized in human fetal spinal cords.Our data show that LIF combines synergistically with high-density reaggregate cultures to support the organotypic reorganization of neurospheres,characterized by surface BMPRⅡ^(+)hNPCs.Our study has provided a new methodology for an in vitro model capable of amplifying human fetal spinal cord cell numbers for>10 passages.Investigations of the role BMPRⅡplays in spinal cord development have primarily relied upon mouse and rat models,with interpolations to human development being derived through inference.Because of significant species differences between murine biology and human,including anatomical dissimilarities in central nervous system(CNS)structure,the findings made in murine models cannot be presumed to apply to human spinal cord development.For these reasons,our human in vitro model offers a novel tool to better understand neurodevelopmental pathways,including BMP signaling,as well as spinal cord injury research and testing drug therapies.